compositions are detailed for i
ted in the presence of non-lethal amounts of antibiotic. Non-lethal doses of
antibiotic weaken the cell membrane facilitating migration of fatty acids from the cell,
DETAILED ACTION
Notice of Pre-AIA or AIA Status
1. The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
2. The Preliminary Amendment filed on June 14, 2024, has been received and entered.
3. Applicant’s election of Group I without traverse on July 13, 2026, is acknowledged.
Claim Disposition
4. Claims 16, 18-19 and 23-25 were cancelled. Claim 1-15, 17, 20-22 and 26 are pending and are under examination.
Drawings
5. The drawings filed on filed on March 1, 2024, has been considered by the examiner.
Information Disclosure Statement
6. To date no Information Disclosure Statements have been filed. Applicant is reminded of the duty to disclose.
reducing
inhibition feedback, leading to an increase in bio-products.
Abstract
7. The abstract is objected to for the following informalities:
“The invention relates to [[M]] methods and compositions [[are detailed]] for increasing bio-products in [[cyanobacteria]] Cyanobacteria where the [[cyanobacteria]] Cyanobacteria is incubated in the presence of non-lethal amounts of antibiotic. Non-lethal doses of antibiotic weaken the cell membrane facilitating migration of fatty acids from the cell, reducing inhibition feedback, leading to an increase in bio-products”.
Appropriate correction is required.
Specification Objection
8. The specification is objected to for the following informalities:
The title of the invention is not descriptive. A new title is required that is clearly indicative of the invention to which the claims are directed. The following is suggested: "Method for increasing pigmentation and/or bioproduct in Cyanobacteria".
The specification is objected to because the priority information is missing from the first page.
Appropriate correction is required.
Claim objection
9. Claims 1-15, 17, 20-22 and 26 are objected to for the following informalities:
For clarity and precision of claim language it is suggested that claim 1 is amended to italicize the organism name (see Cyanobacteria). See also claims 2-3, 12-14, 20-21 and 26 with similar language. The dependent claims hereto are also included.
For clarity it is suggested that claims 2-3, 10, 13 and 14 are amended to spell out the organism name.
For clarity and consistency it is suggested that claims 3-11 are amended to recite “of” in lieu of “according to” (i.e, “The method of claim 1…”, see also claim 2 for example).
For clarity it is suggested that claim 7 is amended to delete “includes” and recite ‘comprises’.
For clarity it is suggested that a comma (,) is inserted before the ‘wherein’ clause (i.e., “The method……claim 1, wherein….”) in claims 8-11.
For clarity it is suggested that claim 10 is amended to read, “….12.8 mg/L was obtained on day 6.
For clarity it is suggested that claim 11 is amended to recite “…chlorophyll [[a]] an accumulation….”.
For clarity and precision of claim language it is suggested that claim 12 is amended to read, “….comprising a [[cyanobacteria]] Cyanobacteria……”.
For clarity it is suggested that claim 21 is amended to delete “any”.
For clarity it is suggested that claim 22 is amended to recite, “fatty acid [[method]] methyl ester[[s comprising]] comprises saturated….”.
Appropriate correction is required.
Claim Rejections - 35 USC § 112
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
10. Claims 1-15, 17, 20-22 and 26 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AlA), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
The claimed invention is directed to a method for increasing pigmentation and/or bioproduct production in Cyanobacteria comprising growing cultures of said cyanobacteria in the presence of non-lethal levels of antibiotics. The claim language does not provide the specific antibiotic, the specific Cyanobacteria, the specific dose that is considered non-lethal thus reads on antibiotic resistance, does not provide the specific bioproduct production or a quantitative measure of increase in any pigmentation, therefore, the encompasses a large variable genus and not adequately described. It is noted that dependent claims recite some of the missing information in claims 1 and 12, however, the independent claims need to stand on their own. The claimed composition has not asserted use. The claimed invention is overly broad and not commensurate in scope with the disclosure in the specification. The claimed encompasses a genus of any antibiotic, any bioproduct, any pigmentation, any Cyanobacteria.
The specification fails to provide a representative number of species for the claimed genus to show that applicant was in possession of the claimed genus. A representative number of species means that the species, which are adequately described, are representative of the entire genus.
The written description requirement for a claimed genus may be satisfied through sufficient description of a representative number of species by actual reduction to practice, disclosure of drawings, or by disclosure of relevant identifying characteristics, for example, structure or other physical and/or chemical properties, by
functional characteristics coupled with a known or disclosed correlation between function and structure, or by a combination of such identifying characteristics, sufficient to show the applicant was in possession of the claimed genus. Vas-Cath Inc. v. Mahurkar, 935 F.2d 1555, 1563-64, 19 USPQ2d 1111, 1117 (Fed. Cir. 1991), states that "applicant must convey with reasonable clarity to those skilled in the art that, as of the filing date sought, he or she was in possession of the invention. The invention is, for purposes of the ‘written description’ inquiry, whatever is now claimed" (See page 1117). The specification does not "clearly allow persons of ordinary skill in the art to recognize that [he or she] invented what is claimed" (See Vas-Cath at page 1116). The skilled artisan cannot envision the detailed chemical structure of the encompassed genus, and therefore, conception is not achieved until reduction to practice has occurred, regardless of the complexity or simplicity of the method of isolation. Adequate written description requires more than a mere statement that it is part of the invention and reference to a potential method of isolating it. The compound itself is required. See Fiers v. Revel, 25 USPQ2d 1601 at 1606 (CAFC 1993).
Therefore, for all these reasons the specification lacks adequate written description, and one of skill in the art cannot reasonably conclude that the applicant had possession of the claimed invention at the time the instant application was filed.
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
11. Claims 1-15, 17, 20-22 and 26 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claims 1, 12 and the dependent claims hereto are indefinite for the recitation of ‘increasing and non-lethal levels’ because it has ambiguity in scope; it leaves the ordinary skilled worker guessing what the quantifiable amount of increase is and compared to what reference point and what amounts are deemed lethal, is there a standard and is it the same for all antibiotics. Claims must point out and distinctly claim the invention, not state preferences or particular options.
Claim 7 is indefinite because it has ambiguity with the recitation of “includes” which is open ended (meaning comprising or including but not limited to). This term is ambiguous because it allows for additional elements or steps beyond what is listed.
Claim 21 is indefinite for the recitation of “any claim 12” and claim 12 does not per se recite options.
Claim 22 lacks clear antecedent basis for the recitation of “said fatty acid method esters”.
Claim 26 is indefinite for the recitation of “increased amounts of hexadecenoic acid” because it fails to inform a skilled artisan of the invention’s boundaries with reasonable certainty. The claim does not recite that the increased amount is ‘relative to’ a baseline amount or control.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
12. This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
13. Claims 1-7 and 13-15 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sitther et al. (US 2023/0340502, August 19, 2021) in view of Gichuki et al. (Dissertation, Morgan State University, May 2022).
The claimed invention is directed to a method for increasing pigmentation and/or bioproduct production in Cyanobacteria comprising growing cultures of said cyanobacteria in the presence of non-lethal levels of antibiotics. The claim language does not provide the specific antibiotic, the specific Cyanobacteria, the specific dose that is considered non-lethal thus reads on antibiotic resistance, does not provide the specific bioproduct production or a quantitative measure of increase in any pigmentation, therefore, the art is applied to these broad concepts.
The primary reference discloses F. diplosiphon cultured with ampicillin. The primary reference discloses amplified gene products and pGEM-7Zf vector containing T7 promoter were double digested with EcoR1 and BamHI restriction enzymes (Promega, USA) and purified using Zymo DNA clean and concentrator kit. Inserts were ligated into the vector at the digested restriction sites with T4 DNA ligase (New England BioLabs, USA) and pGEM-7Zf-phr A expression plasmid constructed to overexpress the photolyase gene. The ligated plasmids were transformed into E. coli FB5α competent cells via heat shock at 42° C. for 20 s followed by incubation on ice for 5 min. The transformed cells were plated on Luria Bertoni (LB) agar plates containing 80 mg L.sup.−1 amphicillin and incubated for 16 h at 37° C. Twenty resistant single colonies were randomly selected, transferred to liquid LB medium containing 80 mg L.sup.−1 amphicillin and grown at 37° C. for 16 h. Plasmids were extracted using the Zyppy plasmid miniprep kit (Zymo Research, USA) and the insert confirmed by PCR and Sanger sequencing at paragraph [0025].
The primary reference further disclosed an Electroporation-Mediated Transformation of the phr A Gene in F. diplosiphon and discloses expression plasmid containing the phr A gene was transformed into F. diplosiphon B481-WT according to parameters described by Tabatabai et al. Competent cells (40 μL) were mixed with ligated purified plasmid DNA and electroporated using a GenePulser Xcell with CE module (Bio-Rad, USA) at 200 Ω resistance, 1.0 kV, and 25 μF capacitance. After incubation on ice for 20 min, the transformant was grown in BG11/HEPES liquid medium for 16 h and plated on LB agar containing 80 mg L.sup.−1 amphicillin. To verify insertion of the phr A gene, PCR was performed using gene specific primers as mentioned above in para. [0021] and products visualized on a 1.5% agarose gel. Paragraph [0022] F. diplosiphon strain (B481-WT) obtained from the UTEX algal repository (Austin, USA) was grown in liquid BG-11 medium containing 20 mM HEPES (hereafter termed as BG-11/HEPES) to an exponential growth phase (optical density at 750 nm of ˜0.6).
At paragraph [0035] Evaluation of Photosynthetic pigment levels photosynthetic efficacy of the wildtype and transformant was quantified as a measure of PSII activity and chlorophyll a content, which provides an estimate of the well-being of photosynthetic cells. To allow maximal irradiation and avoid cell shadowing, cultures grown to an OD.sub.750nm of 0.3 were placed in open petri dishes and irradiated in a UV-B crosslinker (Fisher Scientific, USA) for 60 min. Cultures were grown under conditions mentioned in para. [0017] for three days to allow cell recovery and PSII functionality measured after 24, 48, and 72 h using a MINI-PAM (Walz, Effeltrich, Germany) to measure minimal and maximal fluorescence yield (Fo and Fm). Based on these parameters PSII quantum yield (Fv/Fo) was calculated using the equation Fv/Fm=(Fm−Fo)/Fm. In addition, chlorophyll a (chlα) was measured at the excitation of 420 nm and emission of 680 nm using a microplate reader (Agilent BioTek Synergy H1 Hybrid, USA) and photosynthetic efficacy compared. Characterization of Lipids in the Wildtype and Transformant F. diplosiphon Grown under Simulated UV-B Conditions. At paragraph [0036] Lipid profile of the wildtype and transformant exposed to simulated UV-B conditions (Omaykey UV-B lamps) were compared using GC-MS. Cultures adjusted to 0.1 at OD.sub.750 were grown in 5×7×6 containers and exposed to UV-B for 4 h each day for 15 days to simulate the sun's UV-B radiation effects. Three replicated treatments were maintained and OD.sub.750 measured every three days for a period of 12 days and growth rate calculated. Simultaneous lipid extraction and transesterification was performed as described previously by Tabatabai et al. and fatty acid composition analyzed at the Mass Spectrometry Facility at Johns Hopkins University (Baltimore, MD) using the Shimadzu GC17A/QP5050A GC-MS systems (Shimadzu Instruments, USA). Identification of FAMEs was accomplished by comparing each GC/MS mass spectrum to the Lipid Web archived FAME spectra.
Fatty Acid Methyl Ester Composition in UV-B Irradiated and Non-Irradiated Fremyella diplosiphon Strains, B481-ViAnSa and B481-WT
At paragraph [0044]: F. diplosiphon possesses valuable biodiesel qualities, which can maximize biofuel production. Hence, we compared the high-value saturated and unsaturated FAMEs in the transformant to the wildtype strain. Methyl palmitate, the methyl ester of hexadecenoic acid (C16:0), was found to be the most dominant FAME component, accounting for 53.43% and 51.69% in B481-ViAnSa and B481-WT, respectively. Methyl octadecenoate (C18:0), the second abundant FAME, accounted for 30.12% and 33.02% in B481-ViAnSa and B481-WT respectively. This was followed by methyl octadecenoate (C18:1), which accounted for 22% in B481-ViAnSa and 23.02% in B481-WT. Additionally, we detected methyl tetradecanoate (C14:1), methyl hexadecanoate (C16:1), and methyl octadecadienoate (C18:2) in both strains (Table 2).
TABLE-US-00002 TABLE 2 Quantitative composition of fatty acid methyl esters in transesterified lipids of non-irradiated and UVB radiated F. diplosiphon wild type (WT) and transformant (B481-ViAnSa) strains. Nonirradiated UVB Irradiated FAMEs Type B481-ViAnSa B481-WT B481-ViAnSa B481-WT Methyl palmitate (C16:0) 53.43% 51.69% 42.47% 41.74% Methyl octadecanoate (C18:0) 30.12% 33.02% 23.33% 24.18% Methyl octadecanoate (C18:1) .sup. 22% 23.02% 15.99% 17.05% methyl tetradecanoate (C14:1) 5.19% 5.07% 1.18% 2.07% Methyl hexadecanoate (C16:1) 4.76% 4.59% 0.87% 0.81% Methyl octadecadienoate (C18:2) 3.01% 3.13% 0.91% 0.78%
At paragraph [0045]:… UV-B radiation significantly reduced (p<0.05) the percentage of all FAME components, including methyl palmitate, which was reduced by 20.51% and 19.25% in B481-ViAnSa and B481-WT respectively (Table 2). Due to the exposure of cultures to simulated UV-B radiation for 4 continuous hours, a reduction of FAMEs is expected. However, we observed significantly higher (p<0.05) amounts of saturated FAMEs in both strains irradiated with UV-B when compared to the untreated control. The primary reference does not teach use of lactate dehydrogenase activity, however, Gichuki et al. discloses its use in the manner of determination pertaining to the antibiotic. Gichuki et al. detected in all nZVI-treated cultures in a dose-dependent manner. … amended with 80 mg L-1 ampicillin: (a) Escherichia coli … rate of a non-lethal point mutations for UV radiation and methyl … and also utilized a cyanobacteria Fremyella diplosiphon (see abstract and the entire document).
Therefore, it would have been obvious for one of ordinary skill in the art before the effective filing date of the claimed invention to arrive at the claimed invention as a whole because of the combined teaching of the references. One of ordinary skill in the art would be able to modify the primary reference and combine with the teaching of the secondary reference to arrive at the claimed invention. Motivation exists to combine the teaching of the references because they are analogous art. Moreover, the Supreme Court pointed out in KSA, “a patent composed of several elements is not proved obvious merely by demonstrating that each of its elements was, independently, known in the prior art.” KSA, 127 S. Ct. at 17471. The Court thus reasoned that the analysis under 35 U.S.C. 103 "need not seek out precise teachings directed to the specific subject matter of the challenged claim, for a court can take account of the “inferences and creative steps that a person of ordinary skill in the art would employ.” /d. at 1747. The Court further advised that “[a] person of ordinary skill is...a person of ordinary creativity, not an automation.” /d. at 1742. Therefore, the claimed invention was obvious to make and use at the time the invention was made and was prima facie obvious.
Conclusion
14. No claims are presently allowable.
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/HOPE A ROBINSON/Primary Examiner, Art Unit 1652
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